The FXR agonist obeticholic acid inhibits the cancerogenic potential of human cholangiocarcinoma

Cholangiocarcinoma (CCA) is an aggressive cancer with high resistance to chemotherapeutics. CCA is enriched in cancer stem cells, which correlate with aggressiveness and prognosis. FXR, a member of the metabolic nuclear receptor family, is markedly down-regulated in human CCA. Our aim was to evaluate, in primary cultures of human intrahepatic CCA (iCCA), the effects of the FXR agonist obeticholic acid (OCA), a semisynthetic bile acid derivative, on their cancerogenic potential. Primary human iCCA cell cultures were prepared from surgical specimens of mucinous or mixed iCCA subtypes. Increasing concentrations (0–2.5 μM) of OCA were added to culture media and, after 3–10 days, effects on proliferation (MTS assay, cell population doubling time), apoptosis (annexin V-FITC/propidium iodide), cell migration and invasion (wound healing response and Matrigel invasion assay), and cancerogenic potential (spheroid formation, clonogenic assay, colony formation capacity) were evaluated. Results: FXR gene expression was downregulated (RT-qPCR) in iCCA cells vs normal human biliary tree stem cells (p < 0.05) and in mucinous iCCA vs mixed iCCA cells (p < 0.05) but was upregulated by addition of OCA. OCA significantly (p < 0.05) inhibited proliferation of both mucinous and mixed iCCA cells, starting at a concentration as low as 0.05 μM. Also, CDCA (but not UDCA) inhibited cell proliferation, although to a much lower extent than OCA, consistent with its different affinity for FXR. OCA significantly induced apoptosis of both iCCA subtypes and decreased their in vitro cancerogenic potential, as evaluated by impairment of colony and spheroid formation capacity and delayed wound healing and Matrigel invasion. In general, these effects were more evident in mixed than mucinous iCCA cells. When tested together with Gemcitabine and Cisplatin, OCA potentiated the anti-proliferative and pro-apoptotic effects of these chemotherapeutics, but mainly in mixed iCCA cells. OCA abolished the capacity of both mucinous and mixed iCCA cells to form colonies when administered together with Gemcitabine and Cisplatin. In subcutaneous xenografts of mixed iCCA cells, OCA alone or combined with Gemcitabine or Cisplatin markedly reduced the tumor size after 5 weeks of treatment by inducing necrosis of tumor mass and inhibiting cell proliferation. In conclusion, FXR is down-regulated in iCCA cells, and its activation by OCA results in anti-cancerogenic effects against mucinous and mixed iCCA cells, both in vitro and in vivo. The effects of OCA predominated in mixed iCCA cells, consistent with the lower aggressiveness and the higher FXR expression in this CCA subtype. These results, showing the FXR-mediated capacity of OCA to inhibit cholangiocarcinogenesis, represent the basis for testing OCA in clinical trials of CCA patients

Covid-19 seroprevalence among healthcare workers of a large covid-19 hospital in rome reveals strengths and limits of two different serological tests

Healthcare workers are at the forefront against COVID-19, worldwide. Since Fondazione Policlinico Universitario A. Gemelli (FPG) IRCCS was enlisted as a COVID-19 hospital, the healthcare workers deployed to COVID-19 wards were separated from those with limited/no exposure, whereas the administrative staff were designated to work from home. Between 4 June and 3 July 2020, an investigation was conducted to evaluate the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin (IgG) antibodies among the employees of the FPG using point-of-care (POC) and venous blood tests. Sensitivity, specificity, and predictive values were determined with reverse-transcription polymerase chain reaction on nasal/oropharyngeal swabs as the diagnostic gold standard. The participants enrolled amounted to 4777. Seroprevalence was 3.66% using the POC test and 1.19% using the venous blood test, with a significant difference (p &lt; 0.05). The POC test sensitivity and specificity were, respectively, 63.64% (95% confidence interval (CI): 62.20% to 65.04%) and 96.64% (95% CI: 96.05% to 97.13%), while those of the venous blood test were, respectively, 78.79% (95% CI: 77.58% to 79.94%) and 99.36% (95% CI: 99.07% to 99.55%). Among the low-risk populations, the POC test’s predictive values were 58.33% (positive) and 98.23% (negative), whereas those of the venous blood test were 92.86% (positive) and 98.53% (negative). According to our study, these serological tests cannot be a valid alternative to diagnose COVID-19 infection in progress

Vulvo-perineal reconstruction: Medial thigh septo-fascio-cutaneous island flap

From 1996 through 2001, the authors performed 26 vulvo-perineal reconstructions after vulvar cancer resection. A unilateral or bilateral medial thigh septo-fascio-cutaneous island flap was used in all cases. The vascular supply to this flap is guaranteed by the underlying fascial plexus that arises from the convergence of three longitudinal rows of vessels (anterior, medial, and posterior). The flap is fashioned in a triangle shape and incorporates the underlying muscular fascia, sparing the intermuscular septal perforators. It is advanced in a V-Y fashion and is easily adapted and secured to the recipient site. Competent urethral and anal meati are reconstructed with multiple intercalated flaps. Scarring at the donor site is limited and acceptable. The nerve supply guarantees acceptable sensation at the reconstructed area. The medial thigh septo-fascio-cutaneous flap allows a valid one-stage vulvo-perineal reconstruction with a simple surgical execution process after superficial soft tissue resections

Vulvo-perineal reconstruction: medial thigh septo-fascio-cutaneous island flap.

From 1996 through 2001, the authors performed 26 vulvo-perineal reconstructions after vulvar cancer resection. A unilateral or bilateral medial thigh septo-fascio-cutaneous island flap was used in all cases. The vascular supply to this flap is guaranteed by the underlying fascial plexus that arises from the convergence of three longitudinal rows of vessels (anterior, medial, and posterior). The flap is fashioned in a triangle shape and incorporates the underlying muscular fascia, sparing the intermuscular septal perforators. It is advanced in a V-Y fashion and is easily adapted and secured to the recipient site. Competent urethral and anal meati are reconstructed with multiple intercalated flaps. Scarring at the donor site is limited and acceptable. The nerve supply guarantees acceptable sensation at the reconstructed area. The medial thigh septo-fascio-cutaneous flap allows a valid one-stage vulvo-perineal reconstruction with a simple surgical execution process after superficial soft tissue resections

alpha-Tocopherol and beta-carotene serum levels in post-menopausal women treated with transdermal estradiol and oral medroxyprogesterone acetate.

Estrogens exert a protective effect against atherosclerosis. It is well known that hormone replacement therapy (HRT) can effectively decrease LDL-cholesterol and increase HDL-cholesterol and Apo-Al serum levels. Some recent studies have suggested that estrogens alone or in association with progestins may exert an antioxidant effect on lipids. Besides sex steroids, also vitamins exert an antioxidant effect on LDL and may preserve the endogenous antioxidants of LDL. The aim of our study was to evaluate whether HRT can improve alpha-tocopherol and beta-carotene serum levels in post-menopausal women. Fifteen postmenopausal women with climacteric symptoms were treated with 50 micrograms/24 h estradiol transdermally applied twice a week for 21 days. A daily dose of 10 mg oral medroxyprogesterone acetate was added for 12 days in each treatment cycle. This therapy lasted 6 months. A significant reduction was found in total cholesterol and LDL-cholesterol after treatment. Besides, our study has shown that alpha-toc/LDL and beta-car/LDL ratios significatively increased after treatment, while alpha-tocopherol and beta-carotene serum levels did not change significantly after therapy. These preliminary findings suggest that HRT can preserve the content of alpha-tocopherol and beta-carotene in LDL particles and keep the LDL in a reduced antioxidant state